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. 2023 Jan 14;15(2):517. doi: 10.3390/cancers15020517

Figure 1.

Figure 1

Searching for OTUD7B small molecule inhibitors. (A) A cartoon illustration for the pipeline of searching for OTUD7B small molecule inhibitors. AtomNet was used to search a 4 million small molecule library for hits fitting into the OTUD7B-OTU structure. Top hits were purchased and effects of OTUD7B inhibition by these small molecules were tested by both Akt-pS473 ELISA and Western blotting in A549 cells. Overlapping hits from two assays were identified and tested further. (B) Immunoblot (IB) analyses of whole cell lysates (WCL) derived from control or OTUD7B depleted A549 cells. (C) Representative colony formation analyses images using A549 cells obtained in (B) and quantified in (D). Error bars were calculated as mean+/-SD, n = 3. * p < 0.05 (one-way ANOVA test). (E) A heatmap illustration of Akt-pS473 ELISA results obtained from treating A549 cells with 10 μM of each indicated chemical for 16 h. (F) A list of chemicals with the ability to suppress 30% Akt-pS473 signals from assays in (E). (G) IB analyses of WCL from A549 cells treated with indicated compounds for 16 h. (H) A list of chemicals with the ability to suppress 10% Akt-pS473 signals from assays in (G). (I) A list of overlapped hits from both (F,H).